Flk-1 Antibody Staining Protocol for Immunohistochemistry


Description: Three cell membrane receptor tyrosine kinases, Flt (also designated VEGF-R1), Flk-1 (also designated VEGF-R2) and Flt-4, putatively involved in the growth of endothelial cells, are characterized by the presence of seven immunoglobulin-like sequences in their extracellular domain. These receptors exhibit high degrees of sequence relatedness to each other as well as lesser degrees of relatedness to the class III receptors including CSF-1/Fms, PDGR, SLFR/Kit and Flt-3/Flk-2. Two members of this receptor class, Flt-1 and Flk-1, have been shown to represent high affinity receptors for vascular endothelial growth factors (VEGFs). On the basis of structural similarity to Flt and Flk-1, it has been speculated that Flt-4 might represent a third receptor for either VEGF or a VEGF-related ligand.


Primary Antibody

Name: Flk-1 Antibody

Clone: A-3, Mouse anti-Human

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-6251

Dilution: 1:50 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane
Images: Search image

Additional Information:
Tissue Type: Human colon carcinoma
Fixation: Formalin fixed paraffin sections
Positive Control: Human colon carcinoma
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


Not tested on frozen sections